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Laser Micro-Irradiation to Study DNA Recruitment During S Phase
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Biology
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JoVE Journal Biology
Laser Micro-Irradiation to Study DNA Recruitment During S Phase

Laser Micro-Irradiation to Study DNA Recruitment During S Phase

DOI:
10.3791/62466-v

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07:11 min

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April 16, 2021

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1Department of Biochemistry and Molecular Pharmacology,New York University School of Medicine, 2Laura and Isaac Perlmutter Cancer Center,New York University School of Medicine, 3Howard Hughes Medical Institute,New York University School of Medicine

Chapters

  • 00:04Introduction
  • 00:47Preparation of Cells for Micro-Irradiation
  • 01:18Selecting S Phase Cells for Imaging
  • 02:40Micro-Irradiation for Immunofluorescence Staining or Time Lapse Imaging
  • 03:44Time Lapse Imaging
  • 04:47Results: DNA Recruitment During S Phase After Micro-Irradiation
  • 06:23Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol describes a non-invasive method to efficiently identify S-phase cells for downstream microscopy studies, such as measuring DNA repair protein recruitment by laser micro-irradiation.

Tags

Keywords: Laser Micro-irradiationDNA RepairS PhasePCNASpatiotemporal ResolutionDNA Damage Repair Protein KineticsConfocal ImagingOlaparibEnvironmental ChamberMicroscopeMPlumBinary LinesROIND StimulationTime Schedule

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